Analysis of unsaturated C27 sterols by nuclear magnetic resonance spectroscopy

William K. Wilson, Rhea M. Sumpter, Joshua J. Warren, Peter S. Rogers, Benfang Ruan, George J. Schroepfer

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Abstract

Complete 1H and 13C nuclear magnetic resonance (NMR) signal assignments have been established for 5α-cholestan-3β-ol, 22 unsaturated C27 sterols, and their acetate derivatives. Assignments were made from a combination of 1D and 2D spectra and include stereochemical 1H assignments for the C-22 and C-23 protons of 5α-cholesta-8,24-dien-3β-ol and other Δ24 sterols with a C8 side chain. At the temperature and concentration range described, chemical shifts were generally reproducible to ±0.01 ppm in 13C spectra and ±0.001 ppm in 1H spectra. Except for some overlapped or strongly coupled 1H resonances, chemical shifts are given to these precisions, which are an order of magnitude better than for most data given previously. Full 1H NMR data have been reported previously for only three of the 46 compounds, and 13C data were unavailable for many, including the previously undescribed cholesta-5,8(14)-dien-3β-ol. An extensive set of 1H- 1H coupling constants for the unsaturated sterols indicated considerable conformational diversity, which was confirmed by molecular modeling. The conformational diversity together with other factors led to a complex pattern of 13C substituent-induced chemical shifts (SCS) that appeared to elude any simple empirical correlations with structure. By contrast, the 1H SCS correlated reasonably well with simple structural features. The high precision of the SCS revealed small but measurable effects of a double bond on 1H resonances up to 12 bonds away. Also discussed are the utility and limitations of NMR for the identification of unsaturated sterols, estimation of purity, and analysis of mixtures, with an emphasis on special problems encountered at a microgram level.

Original languageEnglish (US)
Pages (from-to)1529-1555
Number of pages27
JournalJournal of lipid research
Volume37
Issue number7
StatePublished - Jul 1 1996

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Keywords

  • chemical synthesis
  • conformational analysis
  • substituent-induced chemical shifts

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Cell Biology

Cite this

Wilson, W. K., Sumpter, R. M., Warren, J. J., Rogers, P. S., Ruan, B., & Schroepfer, G. J. (1996). Analysis of unsaturated C27 sterols by nuclear magnetic resonance spectroscopy. Journal of lipid research, 37(7), 1529-1555.